Unveiling the positive impact of biofloc culture on Vibrio parahaemolyticus infection of Pacific white shrimp by reducing quorum sensing and virulence gene expression and enhancing immunity.

This study aimed to evaluate and unveil the positive impact of biofloc culture on Vibrio parahaemolyticus infection of Pacific white shrimp by reducing quorum sensing (QS) and virulence gene expression and enhancing shrimp's immunity. The shrimp with an average body weight of 0.50 ± 0.09 g were reared in containers with a volume of 2.5 L, 21 units, and a density of 20 shrimp L-1. The shrimp were cultured for 5 days, with each treatment including biofloc system maintenance with a C/N ratio of 10 and a control treatment without biofloc, followed by a challenge test through immersion using V. parahaemolyticus at densities of 103, 105, and 107 CFU mL-1 initially. The results of the in vitro experiment showed that biofloc suspension can inhibit and disperse biofilm formation, as well as reduce the exo-enzyme activity (amylase, protease, and chitinase) of V. parahaemolyticus. Furthermore, the biofloc treatment significantly reduced the expression of the QS regulatory gene OpaR, the PirB toxin gene, and the virulence factor genes T6SS1 and T6SS2 in both in vitro and in vivo. The biofloc system also increased the expression of shrimp immunity-related genes (LGBP, proPO, SP, and PE) and the survival rate of white shrimp challenged with V. parahaemolyticus.

Immune response, gene expression, and intestinal microbial composition of Pacific white shrimp fed with multispecies synbiotic for the prevention of coinfection disease.

This study aimed to evaluate the application of synbiotic containing multispecies of probiotics with different cell densities in white shrimp rearing against infectious myonecrosis virus (IMNV) and Vibrio parahaemolyticus coinfection. This study used a completely randomized design with five treatments and three replications. One additional replication of each treatment was provided for the lethal sampling. Pacific white shrimp were fed with three dosages of synbiotic multispecies for 30 days, namely 103 CFU mL-1 (Sin 3), 106 CFU mL-1 (Sin 6), and 109 CFU mL-1 (Sin 9), and the controls without synbiotic administration consisted of the positive control (K +) and the negative control (K -). Pacific white shrimp from all treatments, except for the K - , were challenged with IMNV a dose of 100 µL and 106 CFU mL-1 V. parahaemolyticus, injected intramuscularly. Infected Pacific white shrimp showed clinical signs like anorexia, melanosis, empty gut, colorless hepatopancreas, and white necrotic areas in striated skeletal muscles, especially of the distal abdominal segments and uropod. The results showed that administration of synbiotic for 30 days resulted in higher immune parameters, such as total hemocyte count (THC), phenoloxidase activity (PO), respiratory bursts (RB), and total viable bacterial count (TBC) compared to K + /K - . After coinfection, they showed significantly higher levels for THC, PO, RB, gene expression prophenoloxidase (ProPO), and lipopolysaccharide and ß-1.3-glucan-binding protein (LGBP), better clinical signs, and lower mortalities. Sin 9 treatment significantly showed the highest survival rate (SR) compared to the other treatments.

Peptide hydrolysate from fish skin collagen to prevent and treat Aeromonas hydrophila infection in Oreochromis niloticus.

Aquaculture with an intensive system can both increase production and the disease risk. The use of antibiotics to reduce pathogens has bacterial resistance consequence. Peptides have rapidly gained more attention due to their antibacterial and immunomodulatory properties important to combat pathogens. This study aimed to determine the effectiveness of peptide hydrolysate against Aeromonas hydrophila in vitro and in vivo in Nile tilapia. The peptides obtained from fish skin collagen of tilapia, milkfish, and catfish were applied in different doses. Protein content and hydrolysis degree of the peptide were also measured. The peptides were tested in vitro using A. hydrophila. For in vivo test, The peptides and positive control treatments used A. hydrophila bacteria, while negative control treatment used a phosphate buffer saline (PBS). The results showed that all three peptide hydrolysates had similar protein content. The highest hydrolysis degree was shown by milkfish collagen hydrolysate. In vitro assay revealed that 12.5% milkfish skin collagen obtained the highest antibacterial activity. In vivo assay showed that the phagocytic activity, respiratory burst, and lysozyme activity was higher in preventive control than therapy, positive control, or negative control treatments. The growth and survival rate in preventive control was also better than positive control or therapy treatment. This study concludes that the peptide hydrolysate can inhibit the growth of A. hydrophila. Peptide hydrolysate from milkfish skin collagen at 12.5% dose was more effective in preventing the A. hydrophila infection, regarding the immune response, growth, and survival.

Dietary supplementation of Pseudoalteromonas piscicida 1UB and fructooligosaccharide enhance growth performance and protect the whiteleg shrimp (Litopenaeus vannamei) against WSSV and Vibrio harveyi coinfection.

P. piscicida 1Ub and FOS were evaluated for their potential synbiotic effects on growth, immunological responses, and disease resistance against white spot syndrome virus and V. harveyi coinfection, the major pathogen in whiteleg shrimp aquaculture. Four different supplemented diets were used to feed the experimental shrimp for 40 days: control (control, no probiotic, and prebiotic), probiotic (PRO, P. piscisida 1UB 108 CFU mL-1), prebiotic (PRE, FOS 0.5% w/w), and the synbiotic (SYN, PRO + PRE). Shrimp's body weight, weight gain, specific growth rate, feed conversion ratio, survival, digestive enzyme activity, and metabolism-related gene expression were all evaluated on day 40. After 40 days, shrimp were infected with WSSV as the primary infection and V. harveyi as the secondary infection 24 h later. Shrimp were then grown for seven days and fed with a control diet. Survival, total hemocyte count (THC), differential hemocyte, phenol-oxidase (PO), respiratory burst activity (RB), and immune-gene expression were all analyzed at 0, 3, and 7 days after infection. The results showed that the PRO, PRE, and SYN supplementation improves whiteleg shrimp growth performance, immune responses, and protection against WSSV and V. harveyi coinfection. The increased activity of digestive enzymes and metabolism-related genes correlates with higher growth performance. The increase in THC, PO, RB, and immune-related gene expression after coinfection was associated with a significant reduction in shrimp mortality. Our findings also suggest that supplementing with synbiotics improves the overall performance of whiteleg shrimp significantly more than probiotics or prebiotics only.

Growth Performance and Intestinal Microbiota Diversity in Pacific White Shrimp Litopenaeus vannamei Fed with a Probiotic Bacterium, Honey Prebiotic, and Synbiotic.

This study aimed to evaluate the growth performance and intestinal microbiota composition in Pacific white shrimp after probiotic, honey prebiotic, or synbiotic treatment. Pacific white shrimp were treated for 45 days with probiotic (1% (v/w) of Bacillus sp. NP5 RfR probiotic), prebiotic (0.5% (v/w) of honey prebiotic), synbiotic (1% (v/w) of probiotic and 0.5% (v/w) prebiotic), or control (without addition of probiotic and prebiotic). Next-generation sequencing (NGS) was used to assess the effects of these treatments on growth performance and intestinal microbial diversity. The administration of a probiotic, prebiotic, or synbiotic led to increases in specific growth rate, feed conversion ratio, and digestive enzyme activities of amylase, protease, and lipase in Pacific white shrimp. The prebiotic treatment demonstrated the greatest effect, with values of growth rate of 3.09 ± 0.02 (% day-1), feed conversion ratio of 1.45 ± 0.00, and enzyme activities of 1.388 ± 0.0211 IU mg-1 protein for amylase, 0.055 ± 0.0004 IU mg-1 protein for protease, and 0.152 ± 0.0025 IU mg-1 protein for lipase. Analysis of the intestinal microbiota diversity revealed that prebiotic administration caused dominance of the phylum Bacteroidetes, whereas the probiotic and synbiotic treatments caused dominance of the phylum Proteobacteria. Moreover, prebiotic treatment was able to increase the diversity of Microbacterium, Lactobacillus, and Neptunomonas, which are established probiotic candidates in aquaculture. The probiotic, prebiotic, and synbiotic treatments induced a number of operational taxonomic units (OTUs) significantly higher than control treatment, that is, 470, 480, 451, and 344 OTU, respectively.